Objective To study the effect of Mycobacterium tuberculosis (Mtb) Pro-Pro-Glu-59 (PPE59) protein on the biological function of Mycobacterium smegmatis (Ms) and the regulation of host cell immune response. Methods PPE59 gene fragment was obtained by PCR amplification, cloned into pALACE, constructed into recombinant pALACE-PPE59 vector, and electro-transformed into Ms. Western blot was applied to analyse PPE59 expression and subcellular localization. The survival of Ms_Vec and Ms_PPE59 under low acid (pH=3 and pH=5) conditions and active surface pressure sodium dodecyl sulfate (SDS) conditions and their intracellular survival in macrophages were analyzed. ELISA was used to detect the cytokine (IL-1β, IL-6, IL-12, TNF-α and IL-10) expression levels of Ms_Vec and Ms_PPE59 infected macrophages. Results PPE59 protein localized to the cell wall of Ms can enhance the acid-resistance and anti-SDS effect of Ms, which is conducive to the survival of Ms in macrophages. PPE59 significantly decreased the secretion levels of pro-inflammatory cytokines (IL-1β, IL-6, IL-12 and TNF-α), and promoted the secretion levels of anti-inflammatory cytokine (IL-10). Conclusion PPE59 enhances the survival ability of Ms under low acid and SDS pressure and promotes its intracellular survival by regulating the cytokine secretion levels.
Mycobacterium smegmatis/metabolism* ; Macrophages/metabolism* ; Cytokines/metabolism* ; Mycobacterium tuberculosis/metabolism* ; Bacterial Proteins/metabolism* ; Animals ; Mice ; Antigens, Bacterial/metabolism*

Objective To investigate the quality and in vitro release behavior of the sirolimus self-microemulsion-mesoporous silicon sustained release tablets and provide a basis for further research and development of related preparations. Methods The hardness, brittleness and content uniformity of the sustained-release tablets were tested refer to Pharmacopoeia of the People’s Republic of China 2020. Different diameters （10, 11, 12 mm）, different hardness （50, 70, 90 N）, different speed （50, 75, 100 r/min）, different dissolution methods （pulp method, basket method） were investigated. The release conditions of the sustained-release tablets with different pH solution （distilled water solution and 0.4% SDS solution with pH of 1.2, 4.5 and 6.8, respectively） and the in vitro release conditions of the sustained-release tablets were observed. Results The hardness, brittleness and content uniformity of the self-made sustained-release tablets were qualified; different diameters and dissolution methods had no effect on the drug release behavior of the sustained-release tablets in vitro, while the different hardness, different rotational speed and the different pH release media had certain effects. Conclusion The sirolimus self-microemulsion-mesoporous silicon sustained release tablets had good sustained-release effect in vitro and was deserved to further study.

Objective To investigate the relationship between the expression of nuclear factor ⅠB(NFⅠB)in myeloid cells and intestinal inflammation by constructing NFⅠB conditional gene knockout(cKO)mice.Methods Human Protein Atlas database,Genotype-Tissue Expression database,and FANTOM5 database were used to investigate the expression of NFⅠB in inflammatory cells.NFⅠB-floxed mice were constructed using CRISPR/Cas9 technology and hybridized with LyZ2-Cre transgenic mice.Myeloid specific NFⅠB cKO mice(NFⅠBfl/flLyz2-Cre)were obtained by self-crossing the progeny.After the genotype identification of mice by agarose gel electrophoresis,4 NFⅠB cKO mice of C57BL/6N strain were selected as experimental group,and 4 non-cKO mice were selected as control group.Both groups were induced with 2.5％dextran sulfate sodium salt(DSS)under the same condition to establish a chronic colitis model,and the severity of colitis was evaluated by clinical manifestations and histopathology.Results Analysis showed that NFⅠB was expressed in both myeloid granulocytes and monocytes,and the highest expression was found in neutrophils.NFⅠB cKO mice were successfully constructed using CRISPR/Cas9 technology and Cre-loxP system.DSS-induced enteritis NFⅠB cKO mice developed diarrhea,gross blood stools,reduced activity,and weight loss in a short time.The gross examination of the intestines showed that the colon of the NFⅠB cKO mice was significantly shorter than that of the non-cKO mice([8.23±0.35]cm vs[10.30±0.36]cm,P＜0.01).Intestinal H-E staining showed changes in mucosal glandular structure and connective tissue hyperplasia with extensive inflammatory cell infiltration in NFⅠB cKO mice.The histological score of NFⅠB cKO mice was significantly higher than that of non-cKO mice(4.25±0.50 vs 0.50±0.58,P＜0.01).Intestinal immunohistochemical staining showed that more CD11b positive cells were recruited in NFⅠB cKO mice than non-cKO mice.Conclusion Myeloid specific NFⅠB cKO mice have been successfully constructed,and NFⅠB in myeloid cells can reduce infiltration of immune cells(granulocytes or/and monocytes)to inhibit intestinal inflammation.

Objective:To explore the effects of acupuncture combined with Buyang Huanwu Decoction on intestinal flora in cerebral blood flow hypo perfusion model rats with carotid artery stenosis.Methods:Totally 40 rats were randomly divided into sham-operation group, model group, TCM treatment group and acupuncture and drug combination treatment group, with 10 rats in each group. Except the sham-operation group, the other groups were prepared cerebral ischemia model by needle control and thread embolism method. TCM treatment group received Buyang Huanwu Decoction 100 mg/kg for gavage, once a day, and the intervention lasted for 2 weeks. In the acupuncture and drug combination group, based on the TCM treatment group, Baihui and its left and right sides of 2 mm were selected for acupuncture, once a day, and continuous intervention was performed for 2 weeks. Neurological function evaluation and behavioral function score were performed 7 and 14 days after administration, respectively. 16S rRNA sequencing was used to comprehensively characterize the structure and composition of fecal microflora of rats in each group. Linear discriminant analysis Effect Size (LEfSe) was used to analyze the difference of intestinal bacteria among groups.Result:On the 7th and 14th day after administration, compared with the model group, the neurological function score in the TCM treatment group and the acupuncture and drug combination group decreased ( P<0.05), and the behavioral function score increased ( P<0.05). Compared with model group, the Shannon index of TCM treatment group and acupuncture and drug combination group increased ( P<0.05). The abundance of Firmicutes increased ( P<0.05), and the abundance of Bacteroidetes and Proteobacteria decreased ( P<0.05); the abundance of Clostridia increased ( P<0.05), and the abundance of Gammaproteobacteria decreased ( P<0.05). The abundance of Escherichia-Shigella and Bacteroides decreased ( P<0.05); the abundance of lactobacillus significantly increased ( P<0.05). Conclusion:Acupuncture combined with Buyang Huanwu Decoction can improve the symptoms of cerebral hypoperfusion model rats with carotid artery stenosis, and the mechanism may be to increase the abundance of probiotics.

Objective To prepare silymarin phospholipids complex（SM-PC） and investigate its physicochemical properties. Methods On the basis of single-factor tests, the drug-lipid ratio, drug concentration and reaction temperature were selected as the factors of the central composite design and response surface methodology in the preparation of SM-PC by solvent volatilization, and the best process was optimized with the compound rate as the index. And its in vitro dissolution was measured. Results The optimum preparation technology of SM-PC was as follows: acetone was used as compound solvent, the concentration of SM was 8.0 mg/ml, the mass ratio of SM to phospholipid was 1∶1.8, the reaction temperature was 56 ℃ and the recombination rate was（95.15±1.55）% with deviation of less than 3%. The in vitro dissolution test showed that the dissolution of SM-PC was close to 90% in 60 min. The dissolution behavior of main component of silybin was similar to that of silymarin capsules（Legalon ^®）, which was higher than SM-API. Conclusion SM-PC was successfully prepared by central composite design response surface method, which significantly improved the dissolution and laid a foundation for the study of subsequent preparations.

OBJECTIVE To provide reference for strengthening the standardized management of off-label drug use in cancer hospitals. METHODS The evaluation system for off-label drug use was established to standardize the application， approval， and filing process for off-label drug use in our hospital. The changes in off-label drug application quantity， proportion， disease category and drug category in our hospital were compared before （October 1st， 2021-September 30th， 2022） and after （October 1st， 2022- September 30th， 2023） the establishment of the evaluation system； drug items supported by high-level evidence screened by pharmacy department were analyzed statistically. RESULTS The number of off-label drug use applications in our hospital had gradually increased， from 306 pieces in the fourth quarter of 2021 to 3 828 pieces in the third quarter of 2023. In the year before the construction of the evaluation system， there were a total of 4 482 applications for off-label drug use， and in the year after the construction of the evaluation system， there were 11 840 applications for off-label drug use. After the construction of the evaluation system， the proportion of unregistered off-label drug use significantly decreased， compared to the same period last year （P＜0.05）. Among them， there were no unregistered applications for off-label drug use for digestive system tumors， head and neck tumors， and radioactive drugs； lymphoma， breast tumors，urogenital system tumors， cytotoxic drugs and new anti-tumor drugs all had a decrease of over 70% in unregistered off-label drug applications. Twenty-seven off-label drug use items related to 19 drugs supported by high-level evidence were screened by the pharmacy department of our hospital， among which 25 items were drug use beyond indication. CONCLUSIONS The establishment of off-label drug use evaluation system in cancer hospital is helpful to the rational use and refined management of clinical anti-tumor drugs.

Objective:To explore the clinical efficacy of 125I seeds implantation in treating cancer-induced pain and motor dysfunction caused by brachial plexus compression through neurophysiological monitoring. Methods:A retrospective study was conducted on 8 patients (4 males, 4 females; age 58-63 years) who underwent 125I seeds therapy for cancer-induced brachial plexus injury at Hebei Provincial People′s Hospital from January 2021 to August 2023. Pain severity was assessed by using the numerical rating scale (NRS) and motor function was evaluated by using the Fugl-Meyer (F-M) assessment. Electrophysiological monitoring was used to assess changes in sensory and motor branch conduction velocity (CV) of the musculocutaneous nerve, axillary nerve, median nerve, ulnar nerve, and radial nerve before and 3 months after treatment. Paired t-test was used for data analysis. Results:All 8 patients had moderate to severe pain (6 had motor dysfunction). The preoperative and postoperative NRS scores was 5.9±1.0 and 3.3±1.7, respectively ( t=4.93, P=0.002), while F-M scores was 44.8±7.6 and 54.8±5.7, respectively ( t=-3.52, P=0.017). Electrophysiological results showed that 7 patients had lesion involvement in the lower trunk of the brachial plexus, and 1 patient had involvement in the upper trunk. The preoperative and postoperative motor branch CV of the ulnar nerve was (47.2±2.6) and (59.7±8.2) m/s, respectively ( t=-3.17, P=0.034), while the sensory branch CV was (41.8±1.2) and (56.0±5.7) m/s, respectively ( t=-5.82, P=0.001). The nerve CV increased compared to the preoperative ones. Conclusions:125I seeds implantation has good clinical efficacy in treating cancer-related pain and motor dysfunction caused by brachial plexus compression. Changes in electrophysiology can quantitatively monitor the recovery of sensory and motor functions of the brachial plexus.

Objective To introduce and translate the fear of falling avoidance behaviour questionnaire(FFABQ)into a Chinese version and test its reliability and validity.Methods The Brislin translation model was employed to translate and back-translate the original FFABQ.The back-translated FFABQ was modified based on expert consultations and a pre-survey,followed by cultural adaptation evaluation.To assess the reliability and validity of the Chinese version FFABQ(FFABQ-CHN),a convenient sampling method was used to select a total of 269 elderly patients from a general hospital in Xi'an to participate the survey.The item analysis,content validity,exploratory factor analysis,Cronbach α coefficient,split-half reliability,test-retest reliability and paired t-test were employed in the evaluation of the reliability and validity of FFABQ-CHN.Results The Chinese version FFABQ comprised 3 dimensions of walking,daily activities and work/leisure activities,with 14 items in total.A total of 254 patients had responded to the survey.Item analysis revealed that CR values of the 14 items ranged from 3.978 to 11.755(all P<0.05),with correlation coefficients between each item and the total score of the questionnaire from 0.473 to 0.930(all P<0.001).The content validity index at the item level ranged from 0.834 to 1.000,while the content validity index at the scale level was 0.968.Exploratory factor analysis extracted three common factors,with a cumulative variance contribution rate of 85.347%.The Cronbach α coefficient,half-half reliability and test-retest reliability of the questionnaire were 0.929,0.877,and 0.954,respectively.Paired t-test indicated no significant difference between the scores(all P>0.05).Conclusions The Chinese version FFABQ demonstrates strong a reliability and validity in the assessment of activity restriction related to fear of falling in the elderly.Thus,it serves as an effective tool for medical practitioners in evaluation of the restrictions in fear-of-falling-related activities of the elderly.

Objective:To investigate the efficacy and safety of intravenous thrombolysis (IVT) in patients with acute ischemic stroke (AIS) who are currently using direct oral anticoagulants (DOACs).Methods:Patients with AIS admitted to the Stroke Center of Zhengzhou People's Hospital from January 2021 to December 2024 and had taken DOACs within 48 hours prior to onset were included retrospectively. The demographic characteristics, vascular risk factors, laboratory test results, baseline National Institutes of Health Stroke Scale (NIHSS) scores, and other clinical data were collected. The main outcome measure was the functional outcome evaluated using the modified Rankin Scale at 90 days after onset, with a score of 0-2 defined as good outcome. The secondary outcome measures were symptomatic intracranial hemorrhage (sICH) and any bleeding in any location occurring within 36 hours after onset. Multivariate logistic regression analysis was used to determine the independent influencing factors of the outcome. Results:A total of 153 patients were included, with 87 males (56.9%), aged (71.261±6.983) years. Seventy-four patients (48.4%) underwent IVT, and 59 (38.6%) had poor outcome. The good outcome rate in the IVT group was significantly higher than that in the non-IVT group (71.6% vs. 51.9%; χ2=6.274, P=0.012), but there was no significant difference in the incidence of sICH and any bleeding in any location. The baseline NIHSS score of the good outcome group was significantly lower than that of the poor outcome group (8.817±3.677 vs. 11.203±5.060; t=3.361, P<0.001), and the proportion of IVT was significantly higher than that of the poor outcome group (56.4% vs. 35.6%; χ2=6.274, P=0.012). Multivariate logistic regression analysis showed that previous history of stroke or transient ischemic attack (odds ratio [ OR] 3.964, 95% confidence interval [ CI] 1.611-9.753; P=0.003) and high baseline NIHSS score ( OR 1.129, 95% CI 1.034-1.233; P=0.007) were independently associated with the poor outcome, while IVT was independently associated with the good outcome ( OR 0.166, 95% CI 0.068-0.410; P<0.001). Conclusion:For patients with AIS who had taken DOACs within 48 hours before onset, IVT can significantly improve the outcome without increasing the risk of sICH.

Objective:To explore the risk factors affecting the incidence of acute kidney injury(AKI)after liver transplantation(LT).Methods:From November 2019 to November 2022, clinical data were retrospectively reviewed for 105 recipients of classic orthotopic LT.There are 89 males and 16 females with an age range of(50.52±10.35)years.They are assigned into two groups of AKI(66 cases)and non-AKI(39 cases)according to the AKI diagnostic and staging criteria of Global Kidney Disease Prognosis Organization in 2012.General profiles and clinical data(e.g.previous medical history, MELD score, total bilirubin, albumin, serum creatinine level, coagulation function, anhepatic phase and time to surgery)of two groups of recipients are compared.The factors with statistically significant differences are included into multivariate Logistic regression analysis for obtaining independent risk factors for early AKI post-LT.Results:Among them, 66 patients developed AKI within 7 days post-operation with an incidence rate of 62.86%(66/105).The clinical stages of AKI are Ⅰ(46 cases, 69.70%), Ⅱ(10 cases, 15.15%)and Ⅲ(10 cases, 15.15%).Statistically significant inter-group differences exists in age, abdominal surgery history, preoperative serum level of creatinine, operative duration, anhepatic phase and intraoperative plasma transfusion(all P<0.05).Multivariate Logistic regression analysis indicated that abdominal surgery history( OR=5.803, 95% CI: 1.008~33.401, P=0.049), anhepatic phase( OR=1.054, 95% CI: 1.008~1.101, P=0.020)and preoperative serum level of creatinine( OR=0.968, 95% CI: 0.943~0.994, P=0.016)are independent risk factors for early AKI after classical orthotopic LT recipients. Conclusions:Abdominal surgery history, anhepatic phase, and preoperative serum level of creatinine are independent risk factors for early AKI in classic orthotopic LT recipients.